Copper homeostasis in Salmonella enterica: cloning, expression, and purification of CopA transporter and CueO cuproprotein

Marsh, Jeffrey

Student Work

Copper homeostasis in Salmonella enterica: cloning, expression, and purification of CopA transporter and CueO cuproprotein

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Salmonella enterica is an important and frequent cause of gastroenteritis and systemic infections, including those of economic relevance. Salmonella possesses a highly controlled system of transcription regulators, chaperones, membrane transporters, and target cuproproteins that tightly control the distribution of copper across its compartments. Cu+ is required as a redox cofactor in the catalytic centers of enzymes, but free Cu+ is highly reactive and deleterious to cells. Cu+, along with the oxidative burst, is central in host-pathogen interactions as part of the innate immune response. The goal of this major qualifying project is to contribute to understanding the mechanisms of Cu+ homeostasis in the cell envelope of the pathogen Salmonella enterica. Here, the proteins CopA, an inner membrane Cu+ efflux transporter, and CueO, a multicopper oxidase, which are essential in Cu homeostasis, were cloned and purified from Salmonella enterica. Results indicate that CopA and CueO were successfully cloned and expressed in E. coli. Future research should purify these proteins and determine their copper binding kinetics to help understand how Cu+ is transported and stored in Salmonella under conditions of cellular equilibrium.

This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

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